来自钙敏感光蛋白水母发光蛋白的蓝色荧光蛋白是一种耐热酶，催化腔肠素的氧化。
Blue fluorescent protein from the calcium-sensitive photoprotein aequorin is a heat resistant enzyme, catalyzing the oxidation of coelenterazine.
作者信息
Inouye Satoshi
机构信息
Yokohama Research Center, Chisso Corporation, 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan.
出版信息
FEBS Lett. 2004 Nov 5;577(1-2):105-10. doi: 10.1016/j.febslet.2004.09.078.
Blue fluorescent protein from the calcium-sensitive photoprotein aequorin (BFP-aq) was prepared and determined to be a heat resistant enzyme, catalyzing the luminescent oxidation of coelenterazine (luciferin) with molecular oxygen as a general luciferase. After treatment with excess ethylenediaminetetraacetic acid to remove Ca2+ from BFP-aq, the blue fluorescence shifted to a greenish fluorescence. This greenish fluorescent protein (gFP-aq) was identified as a non-covalent complex of apoaequorin with coelenteramide (oxyluciferin) in a molar ratio of 1:1. By incubation with coelenterazine in the absence of reducing reagents, gFP-aq was converted to aequorin at 25 degrees C. BFP-aq and gFP-aq possessing both fluorescence and luminescence activities may work as novel reporter proteins.
制备了来自钙敏感光蛋白水母发光蛋白的蓝色荧光蛋白（BFP-aq），并确定其为一种耐热酶，作为一般的荧光素酶催化腔肠素（荧光素）与分子氧的发光氧化反应。用过量的乙二胺四乙酸处理以从BFP-aq中去除Ca2+后，蓝色荧光转变为绿色荧光。这种绿色荧光蛋白（gFP-aq）被鉴定为脱辅基水母发光蛋白与腔肠酰胺（氧化荧光素）的摩尔比为1:1的非共价复合物。在不存在还原剂的情况下与腔肠素一起孵育，gFP-aq在25℃下转化为水母发光蛋白。具有荧光和发光活性的BFP-aq和gFP-aq可能作为新型报告蛋白发挥作用。
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